摘要
为了阐明TetR家族转录因子SPA7074影响密旋链霉菌Act12的具体机制,利用超高效液相色谱-质谱联用的非靶向代谢组学方法,检测了SPA7074缺失突变株(Δspa7074)与野生型Act12之间的胞内代谢物图谱,并进行多元筛选统计分析。结果表明:在正离子模式下Δspa7074中相较于野生型显著差异代谢物共有55种,为氨基酸及其衍生物、核黄素、S腺苷甲硫氨酸和低碳有机酸等物质,这些化合物多为寡霉素等大环内酯类抗生素生物合成的前体或中间产物,以及能量代谢相关物质;通过胞内相关的次级代谢中间产物的结构分析,对寡霉素的生物合成途径进行了初步的阐述。以上结果说明SPA7074敲除后胞内寡霉素生物的合成显著提高并及时转运出胞外,消耗了大量的氨基酸代谢相关产物,增加了细胞的能量供应,改变了胞内的代谢流。
To elucidate the specific mechanism by which the TetR-family transcription factor SPA7074 affects the Streptomyces pactum Act12.a non-targeted metabolomic approach using ultra-high-performance liquid chromatography-tandem mass spectrometry was used to detect the intracellular metabolite profiles between the SPA7074 deletion mutant(Δspa7074)and the wild-type Act12,followed by multivariate screening statistical analysis.Ultimately,55 metabolites were found to be significantly differentially expressed inΔspa7074 compared to the wild-type in positive ion mode,including amino acids and their derivatives,riboflavin,S-adenosylmethionine,and low-carbon organic acids,which are mostly precursors or intermediates of oligomycin-like macrolide antibiotics biosynthesis,as well as energy metabolism-related substances.The structural analysis of the intermediate products of secondary metabolites related to intracellular biosynthesis pathway of oligomycin D was conducted,which provided a preliminary description of the biosynthesis pathway of oligomycin D.These results indicate that knocking out of SPA7074 significantly increased the intracellular biosynthesis of oligomycin and timely transported it out of the cell,consuming a large amount of amino acid metabolism-related products,increasing the energy supply of cells,and changing the metabolic flow in cells.
作者
郝鹏泽
赵康康
张媛
陈欢
薛泉宏
贾良辉
颜华
HAO Pengze;ZHAO Kangkang;ZHANG Yuan;CHEN Huan;XU Quanhong;JIA Lianghui;YAN Hua(Collage of Life Science,Northwest A&F University,Yangling Shaanxi 712100,China)
出处
《西北农业学报》
CAS
CSCD
北大核心
2024年第11期2156-2162,共7页
Acta Agriculturae Boreali-occidentalis Sinica
基金
陕西省自然科学基础研究计划(2021JM-105,2021JM-087)
国家基金委青年科技基金项目(316001700)。
关键词
密旋链霉菌
非靶向代谢组学
寡霉素D
Streptomyces pactum
Non-targeted metabolomics
Oligomycin D
